کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
11028321 | 1662591 | 2018 | 14 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Assessment of DNA degradation of buccal cells under humid conditions and DNA repair by DOP-PCR using locked nucleic acids
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موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
We analyzed the degradation level of DNA from buccal cells under humid conditions using quantitative PCR analysis. Gauze samples with buccal cells were incubated for up to 12â¯months under three different conditions (25â¯Â°C/dry, 25â¯Â°C/humid, or 40â¯Â°C/humid). The degradation was evaluated based on two degradation ratios (129:41 and 305:41â¯bp). DNA degraded slowly under the 25â¯Â°C/humid condition, and significant differences in the two degradation ratios were detected between 25â¯Â°C/dry and 25â¯Â°C/humid conditions after 12â¯months. Moreover, the degradation rapidly progressed under the 40â¯Â°C/humid condition, and the two degradation ratios in this condition were much lower than those from 25â¯Â°C/dry and 25â¯Â°C/humid conditions after a short incubation period (3â¯months). To evaluate the effect of DNA repair on low-copy degraded DNA, degenerate oligonucleotide-primed PCR (DOP-PCR) was performed before short tandem repeats (STR) genotyping. As a standard DOP-PCR, we used a 22-base primer with 10 degenerate sequences (5â²-CTCGAGNNNNNNNNNNATGTGG-3â²), and additionally designed DOP-PCR primers with 2, 4, 6, or 8 locked nucleic acids (LNAs). When slightly degraded DNA (305:41-bp ratioâ¯=â¯0.60) was used, DOP-PCR significantly increased the fluorescent intensity and success rate of genotyping using Identifiler and Globalfiler kits. In particular, the reaction with four LNAs produced the highest value. However, such benefits were not observed in the analysis of moderately degraded DNA (305:41-bp ratioâ¯=â¯0.13). Although the recovery rates of STR profiles by DOP-PCR were dependent on the degradation level of low-copy DNA, the effectiveness of DOP-PCR highlights the potential of LNA for degenerate sequences.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Legal Medicine - Volume 35, November 2018, Pages 29-33
Journal: Legal Medicine - Volume 35, November 2018, Pages 29-33
نویسندگان
Masaru Asari, Hiroaki Matsuura, Shotaro Isozaki, Chisato Hoshina, Katsuhiro Okuda, Hiroki Tanaka, Kie Horioka, Hiroshi Shiono, Keiko Shimizu,